title: Hematopoietic differentiation and production of mature myeloid cells from human pluripotent stem cells.
abstract: In this paper, we describe a protocol for hematopoietic differentiation of human pluripotent stem cells (hPSCs) and generation of mature myeloid cells from hPSCs through expansion and differentiation of hPSC-derived lin(-)CD34(+)CD43(+)CD45(+) multipotent progenitors. The protocol comprises three major steps: (i) induction of hematopoietic differentiation by coculture of hPSCs with OP9 bone marrow stromal cells; (ii) short-term expansion of multipotent myeloid progenitors with a high dose of granulocyte-macrophage colony-stimulating factor; and (iii) directed differentiation of myeloid progenitors into neutrophils, eosinophils, dendritic cells, Langerhans cells, macrophages and osteoclasts. The generation of multipotent hematopoietic progenitors from hPSCs requires 9 d of culture and an additional 2 d to expand myeloid progenitors. Differentiation of myeloid progenitors into mature myeloid cells requires an additional 5-19 d of culture with cytokines, depending on the cell type.
authors.affiliation: National Primate Research Center, University of Wisconsin Graduate School, Madison, Wisconsin, USA.
authors.forename: Kyung-Dal
authors.initials: KD
authors.lastname: Choi
authors.forename: Maxim
authors.initials: M
authors.lastname: Vodyanik
authors.forename: Igor I
authors.initials: II
authors.lastname: Slukvin
mesh_terms: D002454:Cell Differentiation; D018920:Coculture Techniques / Q000379:methods; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000378:metabolism; D006410:Hematopoiesis / Q000502:physiology; D006801:Humans; D023461:Myeloid Progenitor Cells / Q000166:cytology / Q000378:metabolism*; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D017154:Stromal Cells / Q000166:cytology / Q000378:metabolism*; D013997:Time Factors
chemical_list: D016178:Granulocyte-Macrophage Colony-Stimulating Factor